The bottom of the flow cell was a glass coverslip (24x60mm, Merzel Gläser) coated with 100,000 MW polystyrene (Sigma-Aldrich, 0.5% w/v polystyrene dissolved in toluene and spread onto a clean coverslip). polystyrene beads in deionized MilliQ water were spread over the surface after which it was blown dry with a nitrogen gun. A spacer cut from a double layer of parafilm was placed on the coated side of this coverslip. On top, a clean, uncoated coverslip with two holes for inlet and outlet was placed. Consecutively, the parafilm was melted by heating the flow cell up to 85°C. Anti-digoxigenin antibodies (Roche Diagnostics) dissolved at 100 mg/ml in phosphate-buffered saline (PBS, Sigma-Aldrich) were incubated overnight at 4°C. Lastly, the flow cells were passivated by flushing in Bovine Serum Albumin (BSA) at a concentration of 10mg/ml (New England Biolabs) for at least 1 hour. Buffer exchanges occurred by removing the old buffer at the outlet by a syringe pump (Harvard Apparatus 11 Plus) while simultaneously pipetting new buffer into the inlet.
11 plus
Manufactured by Harvard Apparatus
Sourced in United States
The 11 Plus is a lab equipment product designed for general use in research and scientific applications. It serves as a versatile tool for various laboratory tasks. The core function of the 11 Plus is to provide a reliable and consistent performance for the intended purpose, without further interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Sch23390 hydrocloride, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by New England Biolabs (1 mentions)
Anti digoxigenin antibody, by Roche (1 mentions)
Tecoflex sg 80a, by Lubrizol (1 mentions)
Dm irb, by Leica (1 mentions)
Toluene, by Fujifilm (1 mentions)
Stemi 2000 c, by Zeiss (1 mentions)
Spr 839, by Millar (1 mentions)
Raclopride tartrate salt, by Merck Group (1 mentions)
Synapt g2 hdms, by Waters Corporation (1 mentions)
12 protocols using 11 plus
1
Polystyrene-Coated Glass Coverslip Flow Cell
2
Electrospun TPU-Fibroin Biomaterial Scaffolds
3
Flurothyl-Induced Acute Seizure Protocol
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Flurothyl is a GABAA antagonist that induces acute seizures in rodents and has been used previously by our laboratory and others to investigate the effects of acute seizures on memory (Truitt et al. 1960 ; Prichahd et al. 1969 (link); Holley and Lugo 2016 (link); Holley et al. 2018 (link)). All experimental seizures were induced under a standard fume hood inside a clear acrylic (29 × 16 × 15 cm) inhalation chamber. All mice were allowed to acclimate to the room for 30 min prior to induction. During the procedure, the seizure-designated mouse was placed into a clean transfer cage, and then placed into the acrylic inhalation chamber. Undiluted flurothyl (bis-2,2,2-trifluroethylether), obtained from Sigma-Aldrich (product number: 287571), was pumped into the chamber using an extended glass syringe (14.57 mm) and the Harvard Apparatus model 11 Plus syringe pump at a rate of 30 µL per minute. Flurothyl was allowed to drip at a constant rate onto a platform containing a paper towel strip until the mice exhibited a tonic-clonic seizure as determined by the Racine scale (Racine 1972 (link)). A control mouse was placed in a second acrylic inhalation chamber in parallel with the seizure-designated mouse. After a seizure was induced, both mice were removed from the chambers and placed into individual transfer cages and were monitored for 1 h, allowing time to recover.
4
Electrospinning of Polycaprolactone Scaffolds
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306 mg Purac PCL (PURASORB PC12, Corbion, Amsterdam, Netherlands) was dissolved in 800 µL dichloromethane. 200 µL ethanol was added to this solution and stirred for at least 5 min. The polymer solution was transferred into a 1 mL syringe which was placed on a syringe pump (11Plus, Harvard Apparatus, Holliston, MA, USA). Solution electrospinning was performed with a blunt 27 G stainless steel cannula and a pumping speed of 0.5 mL/h. A high voltage of 12 kV (PS 2403D power supply, Voltcraft, Conrad, Hirschau, Germany) was applied at the spinneret tip. The charged threads of the polymer solution were then deposited on a rotating, grounded collector (Ø 80 mm) perforated with round holes in alternating rows with a hole diameter of 1 mm, a hole distance of 2 mm and a hole depth of 1 mm. After the spinning process, the scaffold was relieved from the collector with water, dried on air, and cut into 50 × 50 mm2 square pieces.
5
RBC Dynamics in Controlled Shear Flow
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Flow experiments were performed as described previously (Dupire et al., 2012 (link); Figure 3A ). Briefly, stored RBCs were diluted (≈500×) in dextran solution and injected in a parallelepiped quartz flow chamber (50 × 10 × 1 mm3) mounted on an inverted microscope (DMIRB, Leica) equipped with a 20× objective and a Photonfocus camera. The fluid was driven by a syringe pump (11 Plus, Harvard Apparatus) at wall shear rates ranging from 1 to 20 s−1 (i.e., shear stresses ranging from 0.04 to 0.8 Pa). RBCs were observed within 50 μm from the bottom wall (zone of constant shear rate) in brightfield microscopy along the direction of the flow gradient (Z-axis in Figure 3A ). Discocyte-shaped RBCs were selected and tracked individually along the flow direction by manually moving the stage to keep the RBC in the center of the field of view. During video acquisition, the shear rate was increased step by step when the flip-flopping RBC displayed a stabilized orbital angle at a given shear rate. Images were recorded at 25 fps and then processed semi-automatically using Matlab routines to measure the orbital angle of the individual RBCs.
6
Quantifying Substrate Concentration in HPLC
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The concentration of a substrate in the eluted solution from HPLC should vary depending on the elution profile. To study the relationship between the signal intensity by corona CAD and the "real" concentration of a substrate, a syringe pump was used to introduce a constant concentration flow of the substrates into the corona CAD. The mother solutions of the substrates (S-03, 04 and 10 -16) were diluted with toluene (Wako Pure Chemical Industries, Ltd., HPLC-grade) in a 10 cm -3 measuring flask to make the sample solutions. The concentration of each sample solution was 50 μg cm -3 . A gastight syringe (Hamilton Co., 81617) filled with these solutions was settled on the syringe pump (Harvard Apparatus, 11plus) and directly connected to the corona CAD. The flow rate of the mobile phase was set to 0.5 cm 3 min -1 . Average signal intensity from corona CAD during a three-minute experiment was used to analyze the effects of the densities. toluene was measured every time between the sample analyses, and their average intensities were subtracted from those of the samples.
7
Surgical Preparation for Left Ventricular Pressure Measurement in Mice
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During surgery, anesthesia was maintained by ventilating with 2% Isoflurane vaporized with 80 ml/min O2. All surgical procedures were performed under magnification (1.5- to 4-fold) via a surgical microscope (Stemi 2000-C, Carl Zeiss). Access to the central venous system was established by cannulation of the left femoral vein (LFV, Figure S1A ).
In all experimental groups, fluid-loss was counteracted by infusing 15 μl/min 0.9% sodium-chloride (NaCl) with a syringe pump (11Plus, Harvard Apparatus). Hereafter, a thoracotomy, as adapted and modified from others (2 (link), 9 (link)), was performed (Figure S1B ). Two cmH2O positive end-expiratory pressure (PEEP) was established in 38 of 48 mice just before diaphragm incision by placing the expiratory ventilation hose 2 cm beneath the water-surface. To validate the established PEEP, on-line airway-pressures were monitored in LabChart 7.3 Pro. After diaphragm incision, a limited costotomy was performed (Figure S1B ). The pericardium was bluntly dissected and a suture (8.0, Suprama) was positioned beneath the inferior caval vein (ICV). Finally, the left ventricular apex was pricked with a 25 gauge needle, and a 1.4-F pressure-conductance catheter (SPR-839, Millar) was inserted (Figure S1C ). Intra-ventricular catheter position was optimized under on-line visualization until rectangular-shaped loops were obtained.
In all experimental groups, fluid-loss was counteracted by infusing 15 μl/min 0.9% sodium-chloride (NaCl) with a syringe pump (11Plus, Harvard Apparatus). Hereafter, a thoracotomy, as adapted and modified from others (2 (link), 9 (link)), was performed (
8
Investigating D1 Receptor Modulation in ALM
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After recovery from the surgery for at least a week, mice were water restricted and trained to perform the cued-licking paradigm. Testing started after 8–12 days of training. 20–40 min before a testing and electrophysiological recording session, mice were head restrained and a 33-gauge inner cannula (0.5 mm projection) was inserted into the guide cannula. A 0.5 μL of solution of either the D1 receptor antagonist (5 μg/μL SCH23390 hydrocloride, Sigma-Aldrich) or sterile saline (0.9%) were infused into ALM at 0.25 μL/min using a syringe pump (11 plus, Harvard Apparatus). Single units were recorded with tetrodes and sorted offline through principal component analysis with offline sorter as described above. Each session of saline infusion was followed, on the day after, by a session with D1 receptor antagonist infusion. Each mouse underwent 1–2 sessions of saline and SCH23390 infusion. In total, we recorded 79 single units from 3 mice in 10 sessions; the average yield for this group was 26.3 neurons per animal and 7.9 neurons per session.
9
Unilateral Infusion of Dopamine Antagonists in ALM
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And 20–40 min before a testing session, mice previously trained in the cued-licking paradigm were briefly anesthetized with 1% isoflurane and a 33-gauge inner cannula (0.5 mm projection) was inserted into the guide cannula. A 0.5 μL of a solution of either the D1 receptor antagonist (5 μg/μL SCH23390 hydrocloride, Sigma-Aldrich, St. Louis, MO), the D2 antagonist (5 μg/μL raclopride tartrate salt, Sigma-Aldrich) or sterile saline (0.9%) was unilaterally infused into ALM at 0.25 μL/min using a syringe pump (11 plus, Harvard Apparatus, Holliston, MA).
10
Binge Ethanol Exposure and Fetal Outcomes
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As we have previously published to model a single binge paradigm, EtOH, at 3g/kg (prepared from 95% Koptec – Decon Labs; Catalog#V1101, delivered in a final volume of 220 µL) or purified water, as a control, was administered by intragastric gavage on GD10 using polyethylene tubing connected to a mini pump (Harvard Apparatus 11 plus, Cat# 70‐2209) set to a flow rate of 100 µl/min (Bake et al., 2012 (link)). Blood (20 µl) was collected from the tail vein using heparinized capillary tubes for the determination of blood EtOH concentrations (BEC) by gas chromatography according to our previous publications (Camarillo & Miranda, 2008 (link); Prock & Miranda, 2007 (link); Santillano et al., 2005 (link)). The average BEC was 112 ± 11 mg/dl. At GD18, pregnancies were terminated, fetuses delivered by laparotomy and measures of fetal weight, crown‐rump length (CRL), midsagittal length (MSL) (O'Leary‐Moore et al., 2010 (link)), biparietal diameter (BPD), fronto‐occipital distance (FOD), and placental efficiency were obtained. Subsequently, tissue was snap‐frozen in liquid nitrogen and stored at −80°C preceding RNA isolation.
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